This study aimed to determine the activity of one Mycoplasma bovis nuclease encoded by MBOV_RS02825 and its association with cytotoxicity. The bioinformatics analysis predicted that it encodes a Ca²⁺-dependent nuclease based on existence of enzymatic sites in a TNASE_3 domain derived from a Staphylococcus aureus thermonuclease (SNc). We cloned and purified the recombinant MbovNase (rMbovNase), and demonstrated its nuclease activity by digesting bovine macrophage linear DNA and RNA, and closed circular plasmid DNA in the presence of 10 mM Ca²⁺ at 22–65 °C. In addition, this MbovNase was localized in membrane and rMbovNase able to degrade DNA matrix of neutrophil extracellular traps (NETs). When incubated with macrophages, rMbovNase bound to and invaded the cells localizing to both the cytoplasm and nuclei. These cells experienced apoptosis and the viability was significantly reduced. The apoptosis was confirmed by activated expression of phosphorylated NF-κB p65 and Bax, and inhibition of Iκβα and Bcl-2. In contrast, rMbovNase^Δ181–342 without TNASE_3 domain exhibited deficiency in all the biological functions. Furthermore, rMbovNase was also demonstrated to be secreted. In conclusion, it is a first report that MbovNase is an active nuclease, both secretory and membrane protein with ability to degrade NETs and induce apoptosis.

Molecules contributing to microbial cytoadhesion are important virulence factors. In Mycoplasma bovis, a minimal bacterium but an important cattle pathogen, binding to host cells is emerging as a complex process involving a broad range of surface-exposed structures. Here, a new cytoadhesin of M. bovis was identified by producing a collection of individual knock-out mutants and evaluating their binding to embryonic bovine lung cells. The cytoadhesive-properties of this surface-exposed protein, which is encoded by Mbov_0503 in strain HB0801, were demonstrated at both the mycoplasma cell and protein levels using confocal microscopy and ELISA. Although Mbov_0503 disruption was only associated in M. bovis with a partial reduction of its binding capacity, this moderate effect was sufficient to affect M. bovis interaction with the host-cell tight junctions, and to reduce the translocation of this mycoplasma across epithelial cell monolayers. Besides demonstrating the capacity of M. bovis to disrupt tight junctions, these results identified novel properties associated with cytoadhesin that might contribute to virulence and host colonization. These findings provide new insights into the complex interplay taking place between wall-less mycoplasmas and the host-cell surface.

This study identified urinary biomarkers for tuberculosis (TB) diagnosis. The urine proteomic profiles of 45 pulmonary tuberculosis patients prior to anti-TB treatment and 45 healthy controls were ...

Lumpy skin disease (LSD) is a severe and highly infectious pox disease of cattle caused by the lumpy skin disease virus (LSDV). To facilitate early control of LSD, this study aimed to develop a new rapid on-site LSDV detection method using an orf068 gene-based recombinase polymerase amplification assay (RPA) coupled with a CRISPR-Cas12a-based fluorescence assay (RPA-Cas12a-fluorescence assay). The results showed that the sensitivity of our RPA-Cas12a-fluorescence assay for detecting LSDV orf068 gene reached 5 copies/μL with plasmid as a template, and 10² TCID₅₀/mL with viral genomic DNA as a template. No cross-reaction with other common bovine viruses was observed. Further, an on-site RPA-Cas12a-fluorescence assay of 40 clinical samples from cattle with or without LSD showed a diagnostic sensitivity of 96.3% (95% CI: 81.0–99.9%) and specificity of 92.31% (95% CI: 62.1–99.6%), which was close to those of the quantitative PCR assay. Therefore, our RPA-Cas12a-fluorescence assay has promising prospects in on-site rapid LSDV detection.

Brucellosis, caused by several species of Brucella, continues to be a significant illness that poses a global threat to public health. China remains a persistent hotspot for brucellosis, despite the implementation of extensive control measures. This study aims to conduct a systematic review and meta-analysis of the seroprevalence of bovine brucellosis in different breeds and regions of China from 2014-2024, and to provide predictions on the future prevalence patterns of brucellosis in cattle and humans. The analysis comprised a total of 80 research studies, which consisted of 187 datasets and a combined sample size of 3,130,706. We estimated the overall pooled seroprevalence of bovine brucellosis in China to be 1.5% (95% CI: 0.6-2.6%). Subgroup analysis revealed that the seroprevalence in dairy cattle was 3.1%, surpassing the seroprevalence in beef cattle (1.3%) and yak (1.5%). Regions that had authorized vaccination programmes exhibited higher seroprevalence (1.8%) compared to regions that did not have vaccination (0.5%). Notably, the study observed a simultaneous rise in both the prevalence of brucellosis in cattle and the number of human brucellosis cases. This suggests that high-quality routine surveillance of brucellosis in cattle will be essential for predicting and responding to cases in humans. Additionally, given the existing prevention and control measures, brucellosis will likely continue to be prevalent in both cattle and people. This systematic review will assist policymakers in adjusting animal surveillance and interregional livestock movement policies, ultimately contributing to the public safety goal of preventing brucellosis in humans by controlling it in animals.

Summary: Mycobacterium tuberculosis ( M. tuberculosis ) regions of difference ( RD ) encode proteins which are potentially useful as diagnostic reagents for tuberculosis ( TB ). In this study, 75 genes from M. tuberculosis RD 1‐ RD 16 were successfully cloned from which 68 proteins were expressed and purified. Three serum pools from patients with pulmonary TB ( PTB ), extra‐pulmonary tuberculosis ( EPTB ) and healthy controls ( HC ) were used to preliminarily screen individual RD proteins. The OD ₆₃₀ ratio of the PTB or EPTB to the HC group ≥ 2‐fold was positive. As a result, 29 proteins were obtained. The serological response to the identified antigens was further verified using 58 PTB samples with 38 sera from smear‐positive PTB ( PTB ‐ SP ) patients and 20 sera from smear‐negative PTB ( PTB ‐ SN ) patients, 16 EPTB samples, 42 latent M. tuberculosis infection samples and 40 HC s by indirect ELISA . With respect to the PTB diagnosis, receiver operating characteristic analysis showed that Rv0222 [area under the curve ( AUC ), 0.8129; 95% confidence interval ( CI ), 0.7280–0.8979] and Rv3403c ( AUC , 0.8537; 95% CI , 0.7779–0.9294) performed better than ESAT 6/ CFP 10 ( AUC , 0.7435; 95% CI , 0.6465–0.8406). Rv0222 and Rv3403c demonstrated the highest diagnostic ability in the PTB ‐ SP group (sensitivity, 86.8%; specificity, 80%), while Rv3403c demonstrated the highest diagnostic ability in the PTB ‐ SN group (sensitivity, 70%; specificity, 80%). With respect to the EPTB diagnosis, Rv0222 exhibited the highest diagnostic value ( AUC , 0.7523; sensitivity, 68.8%; specificity, 87.5%). In addition, the combination of Rv0222 and Rv3403c improved the test for PTB ‐ SN . These results indicate that Rv0222 and Rv3403c would be potential diagnostic biomarkers for active TB serodiagnosis. Mouse experiments demonstrated that Rv0222 and Rv3403c elicited specific cellular and humoral responses which were characterized by production of IFN ‐γ, IgG1, and IgG2a, but a higher level of IgG1 than IgG2a.